Phaseolus germplasm exploration in New Mexico, USA Sep 27 – Oct 8, 2023 (photo: S Dohle, 2023) Population #3400 of wild tepary Phaseolus acutifolius Asa Gray found on Oct. 5, 2023, on the eastern slope of the Peloncillo Mountains, Hidalgo, NM; additional information, see text. 2 Report on a Phaseolus germplasm exploration in New Mexico, USA, Sep 27 – Oct 8, 2023. Participants (alphabetical order): DG Debouck1, S Dohle2, D Marquez3, R Pratt3, M Santaella1, LG Santos1 & M Urban1. 1 International Center for Tropical Agriculture (CIAT), Cali, COLOMBIA 2 United States Department of Agriculture, Pullman, Washington, USA 3 New Mexico State University, Las Cruces, New Mexico, USA Abstract Heat and drought are increasingly reducing bean production in most key areas worldwide. There is thus a justified need to increase variability in genebank collections for two species, P. acutifolius and P. filiformis, well known for tolerance to these stressors. The genebanks of USDA and CIAT currently hold less than five unique accessions of these two species from New Mexico, a state for which there are records since 1851. The compilation of these records in 100 Herbaria indicate 189 populations for six Phaseolus species. An 8-days exploration in four counties (Doña Ana, Grant, Hidalgo and Luna) of SW New Mexico yielded 14 populations, namely nine of P. acutifolius and one of P. filiformis. Nine populations of two species, P. acutifolius and P. grayanus, might be new additions to the flora of that state. Due to late and scarce rains (registered in late August-September 2023, instead of July-August in a regular year), most plants were found in seedling or early flowering-pod filling state which limited our ability to collect mature seed. We obtained soil microbe samples and herbarium specimens were made. Soil samples will be analyzed for all sites where herbarium specimens were obtained. The aberrant climate of the past summer shows the challenge these wild populations face towards drought and early frost patterns, and the capacity of potential survivors to such harsh conditions, which in turn are traits of high interest in bean breeding programs. Follow- up collections have resulted in the procurement of mature P. acutifolius seed from several sites. BACKGROUND AND JUSTIFICATION Over the last fifty years bean production in the Americas has been increasingly pushed towards marginal lands. In the USA (US Dry Bean Council 2023) the shift has been to the western states (primarily North Dakota and Nebraska) from the eastern states (New York and Michigan), in Mexico from the Bajío to Durango, and in Brazil from Sao Paulo to Pernambuco, where drought is a frequent and economically important stressor (Parker et al. 2023). There seems to be a trend for climate to become hotter and drier in western North America (Williams et al. 2020, McKinnon et al. 2021). Climatic alterations and severe drought have caused major disruptions in human societies in A.D. 1100-1400 in northern New Mexico and southern Colorado (Fagan 1999, Diamond 2005). Farmers and agronomists are thus increasingly eager to try high value crops while requiring less water. Tepary beans, Phaseolus acutifolius Asa Gray, originating in Aridoamerica (Nabhan 1985, Garvin & Weeden 1994) have long been known as a drought-hardy crop yielding well in dry areas (Freeman 1912, Waines 1978, Pratt et al. 2023). For New Mexico, Fabian Garcia wrote: 3 “It will be noted that the Tepary produced a larger number of pods and beans than any other varieties. The Ancient Yellow also produced a large number of pods. The New Mexico Pinto was third and the Bayo was last” (Garcia 1917, p. 9). The superiority of tepary over common bean under drought and heat stress conditions was further investigated and demonstrated (Parsons & Howe 1984 and Lin & Markhart III 1996, respectively). In many parts of Africa and the Middle East where the crop has been adopted (Tinsley et al. 1985, Mwale et al. 2020), it is because of its hardiness towards drought. There are thus prospects for breeding tepary per se for arid zones (Small 2014) for autoconsumption on-farm and food processing (e.g. the canning industry of navy beans, seed flour, etc). Another increasingly promising avenue exists for the utilization of tepary germplasm. Wild teparies have been used in interspecific hybridization with common bean to transfer resistance to bruchids (Kuzolwa & Myers 2011) and tolerance to low temperatures (Souter et al. 2017). This is in addition to the early use of tepary as a source of resistance to bacterial blight in common bean (Singh & Miklas 2015). Thus, possibilities of using wild teparies for common bean improvement through wide crossing remain important (Pratt 1983, Thomas et al. 1983, Pratt & Nabhan 1988), considering also the significant synteny between both genomes (Gujaria-Verma et al. 2016, Moghaddam et al. 2021) and the possibilities offered by marker assisted selection in both crops (Assefa et al. 2019). Tepary as a crop has a reduced base of genetic diversity, namely because many landraces went extinct upon the introduction of new irrigation technologies and other hydrological projects particularly over the last century (Nabhan & Felger 1978, Nabhan 2009). Thus, significant genetic diversity of tepary is to be found only in its wild form (with the two variants: var. acutifolius and var. tenuifolius), ranging from the Southwestern USA (Arizona to Trans-Pecos Texas down to Jalisco, Mexico; Freytag & Debouck 2002, Debouck 2019). The presence of wild teparies in New Mexico has been reported previously (Nabhan & Felger 1978, Freytag & Debouck 2002), mostly in the southern counties. In addition, five more species, viz P. angustissimus A. Gray, P. filiformis Benth., P. grayanus Woot. & Standl., P. maculatus Scheele and P. parvulus Greene, have been reported to be present in New Mexico (Freytag & Debouck 2002, Debouck 2021). Further, two species reported from Arizona, viz P. montanus Brandegee and P. ritensis Jones (Debouck 2021), might enter into western New Mexico, thus the inclusion of the Peloncillo Mountains in this exploration. Wild Phaseolus species from New Mexico are poorly represented in genebanks. USDA has one population of wild tepary (a recent collection made by Richard Pratt in Indian Hollow Trail in the Organ Mountains, Doña Ana County). In CIAT, five accessions of wild P. acutifolius (G40075, G40076, G40190, G40191 and G40246) go back to one location at the extreme south of Animas Valley (apparently a population visited several times by Oliver Wendell Norvell between 1948 and 1969) (see also Fig. 4.4. in Dohle et al. 2019), and three accessions of P. angustissimus (G40550, G40685, G40704) referring to the same population collected in the northern Organ Mountains around San Augustine Pass. Bearing these accessions in mind the risk of duplicating material is thus quite low, and with a systematic collecting effort the representation of wild beans can be improved in both USDA and CIAT genebanks. 4 Soil sampling at the very places where plants are growing has been considered, namely in relation to phosphorus, iron and zinc, soil pH and electrical conductivity. These soil-related abiotic factors are important constraints to bean production (Assefa et al. 2019) or affect bean nutritional profile (Beebe 2020). In addition, the microorganisms of the rhizosphere might be very important for the mineral nutrition of the plants, namely nitrogen-related bacteria as Rhizobia or mycorrhizal strains. In this regard, different rhizobia species establishing symbiotic relationships have been reported for the different bean species (Aguilar et al. 2004, Rocha et al. 2020). The findings by Rocha and co-workers (2020) are noteworthy: the nitrogen-fixing symbionts of P. filiformis not only belong to another genus but can withstand temperatures of 40oC and grow under pH of 9.5. Given this and the apparent lack of specific selection of Rhizobia strains by common bean, it seems quite important to study the variability of symbionts associated with wild P. acutifolius and P. filiformis of New Mexico. The primary purpose of this work was thus to collect seeds, namely of wild teparies, to enable others to make progress in breeding different Phaseolus beans. We also wished to collect P. filiformis seed where the opportunity arose. The annual species, P. acutifolius and P. filiformis, are not difficult species in the management of germplasm collections, and thus moderate increase in collections is crucial and feasible. OBJECTIVES • To collect seed for germplasm conservation in USDA and CIAT genebanks. Initial focus on populations of P. acutifolius and P. filiformis, and next of other Phaseolus sensu stricto species (namely from the type localities), and currently not present in genebanks, • To collect and identify microorganisms found in the rhizosphere of such populations, • To collect soil samples very near to the root systems of the collected plants (for herbarium specimens) to perform analysis for soil content in phosphorus, iron and zinc, pH, electrical conductivity and other minerals responsible for salinity. MATERIALS AND METHODS The implements and supplies used in this exploration are listed in the Standard Operations Procedures of CIAT genebank (CIAT 2018). Because two vehicles were used in part of the field work it was found useful to have portable radio walkie-talkie devices. Note the GPS conventional coordinates (obtained from a Garmin GPSmap 62S receiver) are provided for the site where the first plants of the population in reference were found; under the concept of population used in this work variation of 150 m in the three spatial dimensions is possible. The methodology followed was that of crossing altitude gradients through different vegetation types (as explained elsewhere: Debouck 1988) in which populations of Phaseolus have been found in the past (or relatively close) as indicated by herbarium specimens. A 5 compilation of herbarium specimens in the period July 1978 – March 2023 covering 100 Herbaria (86 by personal visits (details in Debouck 2021) and 14 through the SEINet portal) yielded the following previous records (Table 1). Acronyms of Herbaria are reported after Thiers (2023). Table 1 – Numbers of populations by species (verified) and by county of New Mexico obtained from the study of 100 Herbaria in 1978-2023. Counties acut angus filif graya macu parvu Total Hidalgo 13 9 3 25 Luna 6 7 6 1 20 Doña Ana 7 13 3 1 1 25 Eddy 2 2 Grant 3 5 13 17 3 41 Sierra 2 1 4 1 8 Catron 23 10 13 3 49 Socorro 4 1 1 6 Cibola 2 2 Bernalillo 1 1 Valencia 2 2 McKinley 2 2 Sandoval 2 2 Los Alamos 1 1 Rio Arriba 3 3 Total 29 67 9 37 40 7 189 Weather conditions were monitored by periodical checks of the US Drought Monitor for New Mexico (consulted at https://www.droughtmonitor.unl.edu) from June 15 onwards (a likely period for seed germination for the annual or resprouting for the perennial species, if rainfall provides sufficient moisture) to September 14. The Monitor on June 13 indicated absence of drought in western New Mexico, abnormally dry weather in Doña Ana, and severe drought in Eddy and Lea. By September 12, all counties of lower New Mexico were affected by severe drought with extreme conditions in eastern Grant, western Luna, eastern Otero and Eddy and Luna. On the basis of these indications, the counties of Eddy and Lea were not included in the planning for this year. To note, this monitoring came from the question: would it be possible to have isohyets for New Mexico, such as the ones provided by Beck & Haase (1969 p. 14-15), but month after month starting mid-June (thus likely after the last killing frost)? The soil samples were collected as follows: the site very close to the wild bean plant was cleaned and the first 1 cm of debris was removed. Then the soil sample was collected between 3-10 cm in depth into one glass jar. For each site where a population of wild Phaseolus was found, at least 3 jars from different, however closely located, sites were collected. The soil will be air dried, sieved (2-3 mm) and analyzed in the lab of Soil Microbiology and Soil Health, Department of Plant and Environmental Sciences at NMSU. For root collection, the 1-5 root systems were isolated and enclosed in a glass jar with cotton and dried silica gel. 6 From these samples, the Rhizobia strains and other microorganisms of the rhizosphere will be isolated and analyzed in the same lab. Names of places were checked along the following sources: Julyan 1998, DeLorme 2012. Names of plants were checked along the following sources: Dimmit 2000, Dodson 2012. Photographs were taken by DG Debouck unless otherwise indicated. RESULTS a. General: After an 8-days exploration (Annex 1) in four counties of SW New Mexico, a total of fourteen populations were found, as detailed in the next table (Table 2), for four species (out of the six species reported for New Mexico), and a total of 46 herbarium specimens were made and deposited at the NMC Herbarium for conservation and distribution (Annex 2). Soil samples were taken for all populations and deposited at NMSU Las Cruces for further analysis. Table 2 – Basic identifications of the populations found during this exploration. Coll. no. Species County Latitude N Longitude W Elevation m Date 3387 acutifolius Doña Ana 32º 22’ 19.1” 106º 33’ 44.1” 1727 29 Sep 3388 acutifolius Doña Ana 32º 22’ 16.6” 106º 33’ 17.3” 1690 29 Sep 3389 acutifolius Doña Ana 32º 20’ 05.6” 106º 35’ 52.0” 1644 29 Sep 3390 acutifolius Doña Ana 32º 20’ 13.5” 106º 35’ 13.6” 1748 30 Sep 3391 acutifolius Doña Ana 32º 21’ 56.2” 106º 35’ 55.2” 1681 1 Oct 3392 acutifolius Luna 32º 08’ 56.3” 107º 37’ 05.2” 1509 2 Oct 3393 filiformis Luna 32º 08’ 56.3” 107º 37’ 05.2” 1509 2 Oct 3394 grayanus Grant 33º 04’ 13.1” 107º 58’ 56.7” 2308 3 Oct 3395 parvulus Grant 32º 58’ 23.1” 108º 12’ 51.2” 2194 3 Oct 3396 acutifolius Grant 32º 38’ 24.9” 108º 33’ 23.5” 1697 4 Oct 3397 acutifolius Grant 32º 38’ 54.1” 108º 32’ 10.7” 1760 4 Oct 3398 grayanus Grant 32º 37’ 00.4” 108º 24’ 23.0” 1963 4 Oct 3399 grayanus Hidalgo 31º 31’ 04.0” 109º 00’ 10.7” 1673 5 Oct 3400 acutifolius Hidalgo 31º 30’ 56.8” 108º 59’ 11.6” 1653 5 Oct Out of the fourteen populations, nine (#3387, #3388, #3389, #3391, #3392, #3394, #3396, #3397, #3399) could be considered as new additions to the floristic knowledge of New Mexico, as evaluated on the basis of herbarium specimens (or germplasm accessions) compiled for this work. This number should be interpreted along the lack of GPS coordinates on most samples (this technology was not available in old collecting efforts). For example, the specimen EO Wooton 528 (and kept as ARIZ30280; also at NY) was “collected in the Organ Mountains, Dona Ana Co. on Sept 3 1897 at an altitude of 4500 ft.” and could match any (or none) of the populations found this year in the Organ Mountains. The last day (Oct. 6) of this exploration did not yield any population. It was hoped to find remnants of the riparian forest in Leasburg Dam State Park. None of the expected big trees 7 (Juglans, Populus, Salix), as indicated by Dick-Peddie 1993, were found. On the other hand, the altitude gradient from Caballo Reservoir (1280 m) to Emory Pass (2280 m) along State Route 152 in the Gila National Forest was of 1,000 m and gave us six opportunities of survey. The compilation of herbarium records gave for this part of Sierra County P. angustissimus (example: specimen Kenneth Heil 32426 at SJNM) and P. maculatus (example: specimen OB Metcalfe 1104 as CAS282544) as possible. The grazing by cattle on eastern slope of the Black Range and Mimbres Mountains in the Gila National Forest could be considered as severe in some spots (see also Fig. 5 right). b. About species: P. acutifolius Nine populations were found, all of them in stages of early growth, flowering or pod filling, because of late rainfalls, but none with ripe pods and abundant seed production. Some dry pods were found on dried vines of last year (Fig. 6 right), that seemed to be a more regular year in terms of total rainfall and timing as compared to 2023. In this regard, the cover photograph allows this double observation: a carpet of seedlings with eophylls indicating a significant rainfall but until early September 2023 and a good seed production in 2022. This situation allowed for limited seed collecting for now. In view of a follow-up visit in late October, one should note that the weather conditions this year would be particularly challenging for seed production because it is not sure that all plants will produce seed given the probability of early frosts. On the other hand, the seedlings and young plants may offer positive results for sampling the microorganisms associated with their roots (Fig. 6 left). 8 Figure 1 – Wild tepary (upper left, #3396) at early blooming stage in a wash of the western slope of the Big Burro Mountains, note the active pulvini putting leaflets parallel to solar radiation; granitic boulders (upper right, #3387) were a frequent habitat in the Organ Mountains. We posit the boulders may limit access to grazing by cattle. and act as a rainfall ‘catchment’ area to provide added soil moisture at their basal margins, in addition to some shade and shield against the wind. This ‘sheltered’ micro-environment can also be seen for plants of population #3391 in trail between mine house ruin and Hayner Mine adjacent to Rabbit Ears Canyon in the Organ Mountains in Doña Ana County (lower left). Lower right: the arroyo riparian habitat (as defined by Dick-Peddie 1993) of population #3397 on the western slope of the Big Burro Mountains in Grant County. Wild teparies were usually found in open sunny spots in several habitats from scrubland to chaparral vegetation between 1500 and 1760 masl (from 1370 to 1980 masl in the original compilation); Figure 1 (upper and lower right) shows these contrasting environments dependent to topography and moisture availability. Population #3396 displayed the two widths of leaflets, while #3397 and #3400 had the narrow leaflets (the often-mentioned var. 9 tenuifolius). Figure 5 left is noteworthy because it indicates a near altitudinal limit of two species (P. acutifolius and P. grayanus; check p. 23 and p. 243 in Freytag & Debouck 2002) and also the limit between two habitats, one open and often as transition (a step favoring a future domesticate?) and one of climax forest and shaded understory. Follow-up site visits in late October and early November yielded green mature to dry pods from Spring Canyon in the Big Burro Mountains, the La Cueva Recreational area and Sotol Canyon in the Organ Mountains. A hard freeze occurred on 31st October in the Organ Mountains. Plants were observed to be in highly variable condition on 1st and 2nd November. Plants ranged in maturity from juvenile to dry mature and with dry or dehisced leaves to completely green and intact leaves. Some plants bore no pods whereas others bore several in different stages of development. In a few cases, shattered pods were observed. P. filiformis One population (#3393) was found in the Little Florida Mountains in Luna County, from where there was one record (specimen P Knight 2762, kept as UNM46398). That population was found mostly as seedlings germinated this fall because of late rainfalls and as a few dried vines from last year (Figure 2). While the population of wild tepary (#3392) found at the same place was growing between rocks, many seedlings of P. filiformis were growing on open slopes. 10 Figure 2 – P. filiformis (upper left, #3393) as dried vine almost without leaflets from last year (scale bar in cm); upper right, its habitat: the Chihuahuan Desert scrub in Rockhound State Park, Luna County, NM. Lower left: one of the many seedlings germinated because of late rainfalls (note the variegation along main veins of all leaflets and the beginning of heterophylly due to lobing of lateral leaflets of second trifoliolate leaf). Lower right: one of the cacti (Opuntia, cholla group) found in this habitat (lower photos: LG Santos). From existing records for this species, this population was found towards a higher elevation (highest is specimen PJ Alexander 908 collected at 1615 masl); #3393 was found with P. acutifolius (#3392) growing in the same area. The distribution of P. filiformis seems linked to a variant of the Chihuahuan Desert Scrub (Dick-Peddie 1993) although with much less Creosotebush Larrea tridentata. P. grayanus Three populations were found, most of them at vegetative stage, with some plants displaying a few racemes with green and mature pods, giving the opportunity for limited seed collecting for conservation (#3394). The stems sprawling on ground first and twining in upper parts 11 were half the length seen in a normal year; heterophylly was noteworthy as usual in this species. One population (#3394) harbored living adults of bean pod weevils (Apion godmani, identification kindly provided by Dr. Joanie King of NMSU). Because of the tuberous root of this species (Fig. 3 left), the Rhizobia samples were not collected. An appropriate methodology needs to be established before the root can be excavated. Figure 3 – P. grayanus (left, #3394) with a tuberous root (scale bar cm) (photo: M Santaella); right: the pine-juniper forest possibly 2 to 3 years after fire (habitat of #3399) along Geronimo Trail in the eastern part of the Coronado National Forest. Although few, the populations disclosed in this work are inclusive of the altitudinal range (1400-2700 m) of the species in New Mexico. Interestingly, two populations (#3394 and #3399) were found in areas of forest regeneration after massive fire. Resprouting from the tuberous root (that can be the size of a sugarbeet; Fig. 3 left) could be possible after a running fire. P. parvulus One population of this species was found (not far from the type locality: “in deep woods of Pinus ponderosa, in the Pinos Altos Mountains, 14 Aug 1880, EL Greene s.n. (kept as NDG27852”)) at pod maturity, although most plants were small (Fig. 4 left) and without racemes, possibly because of scarce rainfall. 12 Figure 4 – P. parvulus (left, #3395), three plants each from a small tuberous root spherical or fusiform and with a single stem, only the plant in the middle has one raceme with one pod; right, the herbaceous understory with a 2/3 soil cover in the Pinus ponderosa forest, S Dohle indicating a plant of P. parvulus with her left hand (photo: LG Santos). The small plants thrive in the shaded understory of the Ponderosa pine forest (Fig. 4 right). They often don’t show any branching nor any floral structure but leaflets (sometimes lobed) to accumulate photosynthates directly into the small tuberous roots. 13 Figure 5 – Left: the place where as seedlings P. acutifolius #3400 (top) and P. grayanus #3399 (bottom) were found together, on the eastern slope of the Peloncillo Mountains in Coronado National Forest. Right: cattle roaming freely on the western slope of the Big Burro Mountains of Gila National Forest east of Red Rock in Grant County. Figure 6 - Left: M Urban picking a sample of nodules on roots of P. acutifolius #3389, along La Cueva Rocks Trail in the Organ Mountains. Right: M Santaella collecting seeds on dried vines of last year in the same population; the good vine growth is indicative of more favorable weather conditions in 2022 (photos: R Pratt). 14 DISCUSSION AND CONCLUDING REMARKS The afore mentioned findings suggest the following points for discussion. First, while it was a bad year for collecting germplasm as seed, the late germinations of many populations allowed a good sampling of possibly living Rhizobium and other microorganisms. It is not certain that climatic conditions will allow the sampling of seeds towards the end of October 2023, but sampling of microorganisms and soil samples in late September and early October was possible indeed. Variation in amount and distribution of rainfall in summer is almost a characteristic of desert precipitation (Larson & Larson 1997), and collectors have to be flexible. A second point might be related to the accuracy of the climatic forecast (as indicated in the Methods we used the US Drought Monitor 2023). The state of populations of wild teparies disclosed this year indicated that they received enough rain to germinate but until the first fortnight of September, while the model indicated that the drought was getting worse at that time. With the data of 2023 and 2022 (apparently a good year given the number of seedlings and the growth of the vines on bushes), it might be possible to improve accuracy towards finer scales and recent rainfalls. Third, the late and scarce rainfalls of summer 2023 highlight the differences between the survival strategies between species of the genus. The species with tuberous roots such as P. grayanus and P. parvulus (and possibly P. maculatus not found during this exploration) under unfavorable conditions did not invest much in reproductive structures but rather saved photosynthates into their roots. This may also explain why populations of P. grayanus can thrive after running fires. On the other hand, the annual species such as P. acutifolius and P. filiformis seemed to germinate only after heavy showers and entered into early flowering and pod setting. Given the scarcity of rain this year, it seems that many seeds did not germinate and would stay dormant in the soil seed bank until next year. But the strategies followed by the annual might be useful for common bean breeding under terminal drought stress (Rao et al. 2013), namely the fast mobilization of photosynthates towards the sole harvested product, viz. the seeds. In that sense, cover photograph of population #3400 looks like a ‘real-time’ evolutionary experiment (Mayr 2001): under limited moisture and coming frost, only the best genetic combinations will survive through seed production. This may lead to a final consideration: while advances in genomics will enable breeders to re-build a bean crop against these major abiotic stresses (Porch et al. 2013, Parker et al. 2023), the preservation of the remaining wild habitats is critical for our understanding of the physiology of these crop wild relatives in situ, while keeping their potential for further evolution (Dohle et al. 2019), that is, the natural testing of new variants coming up at each generation. Due to an abnormal year our work was useful in pointing to the fragility of these populations while securing some seed for in-depth study and conservation in genebanks and opening up a new horizon in symbionts exposed to the same stresses. ACKNOWLEDGMENTS This field work was made possible thanks to grants generously provided by the United States Department of Agriculture (USDA), New Mexico State University (NMSU) and Centro Internacional de Agricultura Tropical (CIAT). Permits to collect herbarium specimens, seed for germplasm conservation and samples of microorganisms and soil were kindly and swiftly 15 granted by the Forest Service of USDA, the Bureau of Land Management of the Department of Interior and the Land Trust of the State of New Mexico. The staff in these three institutions with whom we were in contact through distance virtual meetings during the preparation phase, and in personal and telephone contact during the collection phase were most supportive. We appreciate very much the nicest welcome of the Dean of the College of Agriculture, Consumer and Environmental Sciences at NMSU, Dr. Rolando Flores. The participants of this exploration warmly thank the interest and support of the following individuals at different steps of this work: Miguel Angel Acosta (CIAT), Esteban Bolaños (CIAT), Efren Delgado (NMSU), Nury Escobar (CIAT), Cristina François (BLM), Lois Grant, Guillermo Hernández (CIAT), Anowar Islam (NMSU), Joanie King (NMSU), Xiufen Li (NMSU), Claudia Maldonado (CIAT), Alexander Medina (CIAT), Maria Mullins (BLM), José Ignacio Ortiz (CIAT), Elizabeth Palzewski (BLM), Gaby Phillips (NMSU), Sarah Fuentes Soriano (NMSU), Carla Olson (WSU), Sam Reiss (BLM), Zachary Rogers (NMSU), Joe Tohme (CIAT), Eliana Urquijo (CIAT), Marilyn Warburton (USDA), and Peter Wenzl (CIAT). The participants also express their gratitude to the Staff of the USDA Plant Exploration Office in Beltsville, namely Dr. Anne Francis. The participants wish to thank the hikers Marta Reece and Melissa Laferrière who kindly provided scouting observations for wild teparies. LITERATURE CITED Aguilar, O.M., O. 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Ernest, C. Estevez de Jensen, F. Gómez, B. Hellier, A.V. Karasev, J.D. Kelly, P. McClean, P. Miklas, J.R. Myers, J.M. Osorno, J.S. Pasche, M.A. Pastor-Corrales, T. Porch, J.R. Steadman, C. Urrea, L. Wallace, C.H. Diepenbrock & P. Gepts. 2023. Genetic resources and breeding priorities in Phaseolus beans: vulnerability, resilience and future challenges. Plant Breeding Reviews 46: 289-420. Parsons, L.R. & T.K. Howe. 1984. Effects of water stress on the water relations of Phaseolus vulgaris and the drought resistant Phaseolus acutifolius. Physiol. Plant. 60 (2): 197-202. Porch, T.G., J.S. Beaver, D.G. Debouck, S. Jackson, J.D. Kelly & H. Dempewolf. 2013. Use of wild relatives and closely related species to adapt common bean to climate change. Agronomy 3: 433- 461. Pratt, R.C. 1983. Gene transfer between tepary and common beans. Desert Plants 5 (1): 57-63. 18 Pratt, R.C., L. Grant, C. Velasco-Cruz & L. Lauriault. 2023. 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New Mexico Chapter. Available at: https://swbiodiversity.org/seinet/collections/index.php. Accessed on April 9, 10, 11, 12, 15 and 16, 2023. Singh, S.P. & P.N. Miklas. 2015. Breeding common bean for resistance to common blight: a review. Crop Sci. 55 (3): 971-984. Small, E. 2014. Tepary bean – an ideal arid zone crop. Biodiversity 15 (2-3): 220-228. Souter, J.R., V. Gurusamy, T.G. Porch & K.E. Bett. 2017. Successful introgression of abiotic stress tolerance from wild tepary bean to common bean. Crop Sci. 57 (3): 1160-1171. Thiers, B.M. 2023. [continuously updated]. Index Herbariorum: A global directory of public herbaria and associated staff. New York Botanical Garden's Virtual Herbarium, New York, USA. http://sweetgum.nybg.org/science/ih/. Accessed on 2 February 2023. Thomas, C.V., R.M. Manshardt & J.G. Waines. 1983. Teparies as a source of useful traits for improving common beans. Desert Plants 5 (1): 43-48. Tinsley, A.M., J.C. Scheerens, J.O. Alegbejo, F.H. Adan, K.C. Krumhar, L.E. 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Science 368: 314-318. 20 Annex 1 – Itinerary of the 8-days Phaseolus germplasm exploration in New Mexico, USA, Sept 27 – Oct 8, 2023. Date Activity Overnight 27-Sep Arrival to El Paso/Las Cruces Las Cruces (Wednesday) 28-Sep Meetings with authorities and principal scientists (NMSU, BLM); Las Cruces (Thursday) final purchase of maps at BLM; visit to NM Univ Press. 29-Sep Exploration in Organ Mountains eastern slope, by taking interstate Las Cruces (Friday) 70 to Organ, pass Organ, after 4 miles take to the left using Aguirre Spring Road; Aguirre Spring Recreation area and Sotol Creek. Then back to Organ and using Baylor Canyon Road to La Cueva Recreation area. 30-Sep Exploration in Organ Mts west, exiting Las Cruces by interstate 10, Las Cruces (Saturday) taking eastwards Dripping Springs Road, then Fillmore Canyon. Then back through interstate 10 and road CR B059 (lower part of Mossman Arroyo) towards Pena Blanca. 1-Oct Exploration in Organ Mts western slope: take interstate 10 SE to El Las Cruces (Sunday) Paso, exiting Las Cruces by Baylor Road, then taking eastwards to Hayner Mine into Rabbit Ears area. 2-Oct Going westward using interstate 10 toward Deming. Exploration in Deming (Monday) City of Rocks State Park, Grant County. Exploration along Spring Trail in Rock Hound State Park, in the Little Florida Mountains. 3-Oct Exploration in the valley of Mimbres, then in the Gila National Lordsburg (Tuesday) Forest, in the Black Range, along the Forest Trail 77 in Cooney Canyon, and later in the vicinity of Redstone east of Scott Peak 4-Oct Exploration of banks of Gila River, N of Red Rock, but gate closed Lordsburg (Wednesday) on Anderson Road. Using Red Rock Road and then Tyrone Thomson Road exploration of western slope of Big Burro Mts, in Gila National Forest. 5-Oct Exploration of one sector of the Peloncillo Mts: Big Creek SW of Lordsburg (Thursday) Animas, taking Cloverdale Road, driving 10 miles south of Animas, then to the west taking Big Creek Ranch Road, but gate closed. Then down to Cloverdale and westward through Forest Road 63 but gate closed. Then using Geronimo Trail eastern slope in Coronado National Forest. 6-Oct (Friday) Lordsburg – Deming –Las Cruces. Exploration of sides of Valley of Las Cruces Rio Grande. Radium Springs (Leasburg Dam State Park), northward along Rio Grande, then westward through State Route 152 to Hillsboro, then Kingston in the Gila National Forest on the eastern slope of Black Range. 7-Oct Closing day at NMSU Las Cruces for the exploration team: check El Paso (Saturday) labeling of herbarium specimens, seeds and soil/ microorganisms, a nd field notes. 8-Oct Departure from El Paso for CIAT and USDA staff. (Sunday) 21 Annex 2 – Labels of the herbarium specimens deposited at the Herbarium NMC of NMSU. United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 29/IX/2023. USA. New Mexico. Doña Ana, Organ Mountains, Aguirre Spring Recreation and Campground area, dry canyon 0.15 km from entrance. GPS: lat. 32o 22’ 19.1”, long. 106o 33´ 44.1”. Elevation: 1727 m. Date: 29 September 2023. Small population of about 20 plants scattered among boulders and granite rocks. Open sunny place, soil gravelly grey, slope of 20-30 degrees. Leaflets healthy with active pulvini. In low height (2-3 m) chaparral of Quercus, Yucca, Macroptilium, and scattered grasses. The wild teparies were found at flowering-green pods state because of a late germination. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, D Marquez, R Pratt, M Santaella, LG Santos & M Urban. No. 3387. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 29/IX/2023. USA. New Mexico. Doña Ana, Organ Mountains, E slope, Sotol Creek. GPS: lat. 32o 22’ 16.6”, long. 106o 33´ 17.3”. Elevation: 1690 m. Date: 29 September 2023. Small population of about 20 plants scattered among granite boulders, on steep slope (30o) of the creek and in sunny open spots. Gravelly coarse brown grey soil, 20% not covered. Plants with 30-100 cm climbing stems, at flowering and green pod stages, leaflets apparently healthy; likely a late germination because of late rainfalls. Chaparral vegetation with Quercus, Juniperus, Dasylirion, Agave, and herbs of Compositae and grasses. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, D Marquez, R Pratt, M Santaella, LG Santos & M Urban. No. 3388. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 29/IX/2023. USA. New Mexico. Doña Ana, Organ Mountains, La Cueva Recreational Area, La Cueva Rocks Trail. GPS: lat. 32o 20’ 05.6”, long. 106o 35´ 52.0”. Elevation: 1644 m. Date: 29 September 2023. Small population of about 10 plants among boulders and rhyolitic rocks, on sunny gentle slope. Soil gravelly grey tan superficial. Plants with climbing stems at flowering stage, wings and standard pink. Associated vegetation: Acacia, Prosopis, Opuntia, Dasylirion, barrell cactus and a few grasses. A bit higher up close to the cliffs a larger population of about 50 plants with dried vines and dry pods from last year, and many 22 seedlings at eophyll stage because of late rainfalls. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, D Marquez, R Pratt, M Santaella, LG Santos & M Urban. No. 3389. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 30/IX/2023. USA. New Mexico. Doña Ana, Organ Mountains, Fillmore Canyon, 1 mi from La Cueva Picnic Area parking lot. GPS: lat. 32o 20’ 13.5”, long. 106o 35´ 13.6”. Elevation: 1748 m. Date: 30 September 2023. A population of 10-20 plants spread along the dry bed of streamlet, in sunny spots. Growing between boulders and andesite rocks; soil gravelly grey brown. Associated vegetation: scrub with Acacia, Allenrolfea, Dasylirion, Fouquieria, Opuntia, Ipomoea and a few grasses. Plants up to 60 cm high in flowering and green pod stage; several seedlings with eophylls and first trifoliolate leaves found due to late germination; dried vines of last year also found. Go up to 1810 masl in the same canyon. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, D Marquez, R Pratt, M Santaella, LG Santos & M Urban. No. 3390. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 1/X/2023. USA. New Mexico. Doña Ana, Organ Mountains, old jeep trail (former Mine House Spring Road, entrance on Baylor Road, 3.7 mi south from intersection with US Route 70) between mine house ruin and Hayner Mine, on W facing slope adjacent to Rabbit Ears Canyon. GPS: lat. 32o 21’ 56.2”, long. 106o 35´ 55.2”. Elevation: 1681 m. Date: 1 October 2023. A population of 20-30 plants scattered on a slope between big boulders and other granitic rocks, in sunny open places. Soil brown stony superficial. Associated vegetation: Yucca, Dasylirion, Opuntia, Quercus, Ipomoea, and a few Compositae. Flowering and green pod stage. Several seedlings found due to late rainfalls, and a few dry vines from last year with open split pods. Go a further 50 m in elevation on the same slope. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, D Marquez, R Pratt, M Santaella, LG Santos & M Urban. No. 3391. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 2/X/2023. 23 USA. New Mexico. Luna, Rockhound State Park, 0.2 km from entrance into Spring Canyon. GPS: lat. 32o 08’ 56.3”, long. 107o 37´ 05.2”. Elevation: 1509 m. Date: 2 October 2023. A population of about 10 plants in a dry wash, in an open sunny space. Two kinds of plant material found: seedlings because of late rainfalls and a few dry vines from last year with open pods. Soil stony superficial organic brown tan, with andesite rocks. Associated vegetation: a few low-height (2.5 m) Quercus and Prosopis, Juniperus, Dasylirion, Opuntia (Cholla), Larrea tridentata, several different annual Compositae and a few grasses already dried. Found together with Phaseolus filiformis (# 3393). Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3392. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus filiformis Bentham, ident. DG Debouck, date 2/X/2023. USA. New Mexico. Luna, Rockhound State Park, 0.2 km from entrance into Spring Canyon. GPS: lat. 32o 08’ 56.3”, long. 107o 37´ 05.2”. Elevation: 1509 m. Date: 2 October 2023. A population of about 50 plants in and on borders of a dry wash, in an open sunny space. Two kinds of plant material found, mostly as seedlings because of late rainfalls and a few dry vines from last year with open pods. Soil stony superficial organic brown tan, with andesite rocks, only about 50% cover. Associated vegetation: a few low-height (2.5 m) Quercus and Prosopis, Juniperus, Dasylirion, Opuntia (Cholla), Larrea tridentata, several different annual Compositae and a few grasses already dried. Found together with Phaseolus acutifolius (# 3392). Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3393. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus grayanus Wooton & Standley, ident. DG Debouck, date 3/X/2023. USA. New Mexico. Grant, Gila National Forest, Mimbres River Trail no. 77, E facing slope of Cooney Canyon. GPS: lat. 33o 04’ 13.1”, long. 107o 58´ 56.7”. Elevation: 2308 m. Date: 3 October 2023. A population of about 20-30 scattered plants on a flat terrain then on a slope, down to 2160 m where the creek, found at pod filling stage with 2-4 seeds per pod. Stems up to 80 cm long sprawling on ground and then twining in upper parts, with marked heterophylly. Plants being part of the understory of a Pinyon pine Juniper (J. deppeana) forest recovering from the Black Fire of 2022, with Quercus, Yucca, Opuntia, Geranium, and a few grasses such as Bouteloua. Soil gravelly brown rich in organic matter, derived from diorite rock, 15% not covered. Slight to steep slope, in semi shady habitat. Grazing by horses in some spots. Some leaves punched by a beetle. Soil samples have been handed over to NMSU for further analysis. 24 Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3394. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus parvulus Greene, ident. DG Debouck, date 3/X/2023. USA. New Mexico. Grant, Gila National Forest, on the eastern slope of Scott Peak, 1 km N of Redstone, 0.15 km off State Route 15 to the west. GPS: lat. 32o 58’ 23.1”, long. 108o 12´ 51.2”. Elevation: 2194 m. Date: 3 October 2023. A small population of 10-20 plants scattered in the understory of Pinus ponderosa forest (15 m plus high), with a few Juniperus, Lupinus, Geranium, a few Compositae and grasses, with 30% of soil not covered. Gentle slope, shady habitat. Soil loamy brown rich in organic matter, possibly deep. Plants of 20-30 cm high, mostly found at vegetative stage or with 1-2 racemes with 1-2 pods near maturity. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3395. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 4/X/2023. USA. New Mexico. Grant, on the W border of Gila National Forest, W slope of the Big Burro Mountains, in the surroundings of Red Rock Road. GPS: lat. 32o 38’ 24.9”, long. 108o 33´ 23.5”. Elevation: 1697 m. Date: 4 October 2023. A population of about 20 plants scattered on the borders of a dry wash, with twining stems 40-60 cm high, at flowering-pod filling stage for the earliest plants. Soil grey tan gravelly and sandy as product of erosion of the slopes, one third not covered, derived from miscellaneous rocks of volcanic origin. Open habitat full sunlight, active pulvini. A low-density alluvial chaparral with Juglans, Quercus and Juniperus on the borders of the wash, Opuntia cholla, Dasylirion, Ipomoea and several Compositae. Foliage healthy. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3396. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus acutifolius A. Gray, ident. DG Debouck, date 4/X/2023. USA. New Mexico. Grant, on the W border of Gila National Forest, W slope of the Big Burro Mountains, in the surroundings of Axle Canyon Road. GPS: lat. 32o 38’ 54.1”, long. 108o 32´ 10.7”. Elevation: 1760 m. Date: 4 October 2023. A population of about 10 plants with very 25 narrow leaflets as compared to # 3396, scattered along a dry wash. Open sunny habitat. Soil brown organic, sandy or gravelly alluvium derived from intrusive rocks and sandstone, with no cover up to 30%. Plants at flowering stage (because of late rainfalls), twining stems 50 cm high. Associated flora: Populus close to streamlet borders, Juniperus and Quercus scattered in the immediate surroundings, Ipomoea, some Desmodieae and Compositae. Horse grazing. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3397. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus grayanus Wooton & Standley, ident. DG Debouck, date 4/X/2023. USA. New Mexico. Grant, on the W border of Gila National Forest, W slope of the Big Burro Mountains, on the western slope of Chocolate Peak, about 2.2 miles east of intersection of Tyrone Thompson Road with State Route 90. GPS: lat. 32o 37’ 00.4”, long. 108o 24´ 23.0”. Elevation: 1963 m. Date: 4 October 2023. A population of about 20 plants, most of them in vegetative stage, with stems 60 cm long sprawling and then twining. Half shady habitat, on slopes and flat parts on sides of a dry wash. Soil brown rich in organic matter, partly baren dried, derived from andesite. Associated flora: Pinus ponderosa, Juniperus, Quercus, Opuntia (cholla), Vitis arizonica, Cologania, Desmodium, Geranium, and several Compositae. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3398. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. Phaseolus grayanus Wooton & Standley, ident. DG Debouck, date 5/X/2023. USA. New Mexico. Hidalgo, Peloncillo Mountains, Coronado National Forest, along Forest Road 63 beyond Field Ranch. GPS: lat. 31o 31’ 04.0”, long. 109o 00´ 10.7”. Elevation: 1673 m. Date: 5 October 2023. A small population of about 25 plants and most in vegetative stage (because of late/ few rainfalls), just a few stems with green pods and one with mature seeds. Growth habit: stems sprawling on the ground. Half shade in pine-juniper forest, recovering after the Foster Fire of 2022 or possibly an earlier fire. Soil loamy brown rich in organic matter, half not covered; mixed intrusive rock in the wash, andesite on the dry slopes of the wash. With Agave, Vitis arizonica. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3399. + + United States Department of Agriculture, Agricultural Research Service, Pullman, WA; New Mexico State University, Las Cruces, NM, USA; International Center for Tropical Agriculture, Palmira, Colombia. 26 Phaseolus acutifolius A. Gray, ident. DG Debouck, date 5/X/2023. USA. New Mexico. Hidalgo, Peloncillo Mountains, Coronado National Forest, along Geronimo Trail 1.2 miles westward beyond the entrance sign of the Coronado National Forest. GPS: lat. 31o 30’ 56.8”, long. 108o 59´ 11.6”. Elevation: 1653 m. Date: 5 October 2023. A population of about 30 and more plants, mostly at seedling stage or three trifoliolate leaves (with very narrow leaflets) because of late rainfalls; in spots inaccessible to cattle a carpet of hundreds of seedlings with eophylls has been noted, also dried twining vines of last year. On sides of a dry wash, in sunny to half shaded spots. Soil brown rich in organic matter derived from puddingstone. In juniper-oak forest following Foster Fire (2022, or earlier), with Vitis arizonica, Geranium, and several Compositae; Phaseolus grayanus (not different from # 3399) at various stages of development including seedlings has been found in the surrounding. Soil samples have been handed over to NMSU for further analysis. Coll. DG Debouck, S Dohle, R Pratt, M Santaella, LG Santos & M Urban. No. 3400.